Composition and method for inhibiting, preventing, or ameliorating complications associated with ingestion of a medicinal, chemical, or biological substance or agent

ABSTRACT

The application relates, in part, to a therapeutic composition comprising a pharmaceutically active agent and a diarrheagenic agent.

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority to and the benefit of U.S. ProvisionalApplication No. 60/691,044, filed on Jun. 16, 2005, the content of whichis hereby incorporated by reference in its entirety.

BACKGROUND

A therapeutic drug can be toxic or lethal if ingested at or above aspecified dosage. A chemical substance or biological agent not intendedfor oral consumption can have dire health or safety consequences ifingested. Worldwide, thousands of accidental or intentional overdosingor poisoning incidents result in significant morbidity and mortalityeach year. Healthcare and other costs stemming from treating patientswho suffer from complications associated with overdosing on medicines oringestion of harmful chemical or biological substances or agents arestaggeringly high.

A medication, a chemical substance, or a biological agent can beingested beyond a safe quantity in any of a variety of contexts. Forexample, and without limitation, a child may ingest a sufficiently largequantity of a medicinal or other chemical or biological substance bymistaking the substance for candy or other food or drink; an adolescentor adult may ingest a harmful, or in some instances lethal, dose of anover-the-counter (OTC) or prescription medication in an attempt tocommit suicide; a patient may inadvertently ingest an inappropriatelyhigh dose of a medication by misunderstanding a physician's, apharmacist's, or a pharmaceutical manufacturer's instructions, or bymistaking one medication (and hence its permitted dosage) for another;and a patient being treated for drug addiction may overdose on, forexample and without limitation, methadone, a synthetic opiate used totreat heroine dependence.

Methods to date have suggested applying to medicinal compositions anemetic coating to induce emesis (vomiting) and expel a toxic substancefrom the stomach. However, it has proven difficult to find an effectiveemetic preparation that has tolerable side effects in a broad patientpopulation. Additionally, emesis carries the risk that a patient may gagduring uncontrollable vomiting, and depending on his or her state ofintoxication—suffer an accelerated death by asphyxiation

Moreover, emesis is unsuitable for preventing potentially toxic or otherharmful effects in certain contexts. This is at least in part becauseemesis essentially fails to expel a substance poised for absorption bythe small or large intestine after the substance has already passedthrough the stomach.

Accordingly there is a need for an improved composition and method toinhibit, prevent, or ameliorate complications associated with ingestion,or typically an excessive ingestion, of a medicinal, chemical, and/orbiological substance or agent. There is also a need for a compositionand method to discourage intentional overdose on a medicinal, chemical,and/or biological substance or agent, for example and withoutlimitation, a therapeutic composition such as a psychoactive drug.

SUMMARY OF THE APPLICATION

Accordingly, the present application provides methods and compositionsto address such needs.

In one aspect, when a pharmaceutically active agent, which is safe wheningested at a recommended level or below a threshold amount, is ingestedby a subject in excess, either by intentional or accidental overdose,and thereby becoming potentially lethal, toxic, or otherwise harmful orundesirable, the compositions and methods described herein reduce thelikelihood of, or in some instances prevent, death, injury, or otherharm by producing diarrhea in the subject ingesting the excesscomposition comprising the pharmaceutically active agent. The subjectcan be a human or an animal, preferably a mammal. In certainembodiments, the diarrhea is rapid and is induced by employing one ormore biological, chemical, or biochemical compounds or agents havingdiarrheagenic properties. In certain embodiments, the diarrheageniccompounds have predictable and thus regulatable effects in a subject.

In certain embodiments, the application provides a therapeuticcomposition comprising a pharmaceutically active agent and a diarrhoeticor diarrheagenic agent. The diarrheagenic agent is a substance or agentcapable of inducing diarrhea, if the therapeutic composition is taken atan undesirable high dose, for example, a dose exceeding a prescribed orotherwise safe or threshold quantity. If a subject takes a prescribeddose or a dose below a threshold amount, however, the pharmaceuticalagent in the therapeutic composition has its proper or intendedpharmacological effect, and the therapeutic composition would not inducediarrhea or other undesired side effects in the subject.

In certain embodiments, the diarrheagenic agent is coated onto thepharmaceutically active agent.

In certain embodiments, when the therapeutic composition is ingested bya subject at an appropriate level (e.g., prescribed dosage or below athreshold level), the cumulative amount of the diarrheagenic agentpresent in the subject is below a threshold level that is required toinduce diarrhea. Accordingly, the pharmaceutically active agent isformulated in the therapeutic composition such that the agent will bereleased in the subject at a desirable level or dosage. When thetherapeutic composition is ingested by a subject at a level exceeding anappropriate level (e.g., prescribed dosage or below a threshold level),the cumulative amount of the diarrheagenic agent present in the subjectis above the threshold level that is required to induce diarrhea andthereby induces diarrhea. In certain embodiments, the diarrhea is rapid,or in certain instances, instant. Accordingly, the pharmaceuticallyactive agent is formulated in the therapeutic composition such that thecumulative amount of the agent released in the subject will be lowerthan the amount of the agent that would have been released in theabsence of the diarrhea.

In certain embodiments, the diarrheagenic agent is a toxin, a viralprotein, a laxative, a ligand for an enterotoxin receptor, any analog ofany of the foregoing, or any agent that is capable of inducing diarrheain a subject, or any combination of the foregoing. Examples of toxinsinclude, such as for example, a bacterial enterotoxin or an analogthereof, e.g., an STa peptide or its analog. An enterotoxin analog canbe a peptidomimetic based on a naturally-occurring enterotoxin or avariant thereof. A naturally-occuring enterotoxin can be a peptidehaving an amino acid sequence of any of SEQ ID NOs:1-11. An enterotoxinpeptide can be a peptide having an amino acid sequence at least 70%,75%, 80%, 85%, 90%, 95%, or 100% identical to any of SEQ ID NOs: 1-11.Accordingly, an enterotoxin peptide can be a naturally-occuringenterotoxin or a variant or mutant thereof. A mutant enterotoxin peptidemay have amino acid mutations at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10positions of any of SEQ ID NOs: 1-11. An amino acid mutation can be asubstitution, a deletion, or an addition of the amino acid at thecorresponding position of the respective amino acid sequence.

Examples of laxatives include, without limitation, aloe vera, bisacodyl,casanthranol, cascara sagrada, castor oil, dehydrocholic acid,phenolphthalein, picosulfate, senna, sennosides, or any combinationthereof.

In certain embodiments, the diarrheagenic agent includes a ligand thatbinds an enterotoxin receptor, such as for example, the human guanylatecyclase-coupled heat stable enterotoxin receptor as described in deSauvage et al., J. Biol. Chem. (1991) 266: 17912-17918. The ligand canbe a small molecule (including a natural product, a synthetic molecule,or a metabolite) a peptide or a peptide analog (e.g., a peptidomimetic),a nucleic acid, an aptamer, a naturally-occurring ligand for therespective receptor, or a synthetic ligand (including mutant or variantof a naturally-occurring peptide ligand).

In certain embodiments, the diarrheagenic agent includes a peptidecomprising guanylin, uroguanylin, or a combination thereof

The pharmaceutically active agent of a therapeutic composition of theapplication can be any drug, biologic, or dietary supplement, or anycombination thereof. In specific embodiments, the pharmaceuticallyactive agent is a compound the overdose (e.g., ingestion by a subject atan inappropriate level or a level exceeding the recommended dosage) ofwhich is harmful (e.g., toxic, injurious, or deadly) to a subject. Thesubject can be a human or animal patient in a general population or in aparticular population. For example, a subject can be a pediatricpatient, a pregnant or nursing woman, an elderly patient, or a patienthaving another condition or disease (e.g., an addiction to thepharmaceutically active agent or a hepatic disease) that makes thepatient more vulnerable to a higher-than-desirable level of the specificpharmaceutically active agent or any pharmaceutically active agent.

In certain embodiments, the therapeutic composition comprises, a secondcoating (e.g., an enteric coating) that substantially envelops the firstcoating, and thereby reduces or inhibits the absorption of thetherapeutic composition and/or the drug, chemical substance and/orbiological agent of the therapeutic composition by the stomach.

Additionally, the enteric coating may inhibits, discourages, or preventsthe diarrhoetic agent from being inactivated by the acidity of theenvironment of the stomach. When the composition reaches the small orlarge intestine, the diarrhoetic coating is available to interact withthe small or large intestine essentially prior to absorption of thedrug, chemical substance and/or biological agent of the composition. Ifa sufficient quantity of the composition (e.g., in the form of one ormore medicinal pills) is ingested, severe and typically involuntarydiarrhea ensues, expelling potentially harmful contents of the pillsfrom the gastrointestinal (GI) tract.

Examples of enteric coatings include, without limitation, a hydrophilicpolymer, an enteric polymer, a pH modifier, or any combination thereof.

In certain embodiments, a therapeutic composition of the presentapplication comprises an agent that would deter tempering or breaking ofthe pharmaceutical composition to cause release of one or morepharmaceutically active agents included therein. Such an agent can be anirritant, such as for example, an irritant added to prevent someone fromcrushing a pill of an extended-release formulation and therebydestroying its extended-release characteristics as described in U.S.Patent Application Publication No. 20030125347.

Accordingly, certain embodiments of the application provides atherapeutic composition containing a medicinal, chemical, and/orbiological substance or agent coated with a diarrhoetic compound; and asecond protective coating to delay, reduce, discourage, or in someinstances prevent, absorption of the therapeutic compound or thediarrhoetic coating in the stomach such that an overdose of the compounddoes not become lethal or otherwise unacceptably harmful.

In one embodiment, if the composition according to the application isingested at an inappropriately high level (e.g., above a thresholdlevel), the diarrhea-inducing compound acts in the small or largeintestine to induce diarrhea, thereby expelling the therapeuticcomposition from the GI tract prior to the therapeutic composition beingabsorbed into the blood stream and producing toxic or other harmfuleffects.

Another aspect of the present application provides a diarrheageniccomposition useful as a pharmaceutical coating, excipient, or carrier.The diarrheagenic composition comprises a diarrheagenic agent, such asfor example, a laxative, a toxin (e.g., a bacterial enterotoxin such asSTa) or a ligand for an enterotoxin receptor (e.g., a human guanylatecyclase-coupled receptor). The diarrheagenic composition can be combinedwith a pharmaceutical composition including one or more pharmaceuticallyor therapeutically active agents, and the composition is useful inpreventing harmful effects, for example, those caused by accidental orintentional overdose of the pharmaceutical composition. In certainembodiments, the diarrheagenic composition may further comprise an agentthat would deter tempering or breaking of the pharmaceutical compositionto cause release of one or more pharmaceutically active agents includedtherein. Such an agent can be an irritant, such as for example, anirritant added to prevent someone from crushing a pill of anextended-release formulation and thereby destroying its extended-releasecharacteristics as described in U.S. Patent Application Publication No.20030125347. In certain embodiments, the diarrheagenic compositionfurther comprises an enteric coating.

Accordingly, the application provides a composition for preventing orreducing potential harm from overdose ingestion, comprising: an amountof a core ingredient below a threshold dose; and a diarrheagenic coatingenclosing the core ingredient and containing an amount of adiarrheagenic substance, the amount of the diarrheagenic substance beingsufficient to cause diarrhea to expel at least a portion of the coreingredient from the gastro-intestinal tract of a specimen if thespecimen ingests the composition approximately at or above the thresholddose, thereby reducing potential harm from overdose ingestion of thecomposition. The composition may further include a controlled-releasecoating containing a controlled-release substance to adjust at least oneof a time and a venue of absorption of the core ingredient in thegastro-intestinal tract, the controlled-release coating interposedbetween, and separating, the core ingredient and the diarrheageniccoating. The composition may further include an enteric coatingcontaining an enteric substance, the enteric coating enclosing thediarrheagenic coating to at least partially reduce premature absorptionof at least one of the core ingredient and the diarrheagenic substancein the gastro-intestinal tract.

The application also provides a composition for preventing or reducingpotential harm from overdose ingestion, comprising a mixture of: anamount of a core ingredient below a harmful threshold dose; and anamount of a diarrheagenic substance, the amount of the diarrheagenicsubstance being sufficient to cause diarrhea to expel at least a portionof the core ingredient from the gastro-intestinal tract of a specimen ifthe specimen ingests the composition approximately at or above thethreshold dose, thereby reducing potential harm from overdose ingestionof the composition. The composition may further include an entericcoating containing an enteric substance, the enteric coating enclosingthe mixture of the core ingredient and the diarrheagenic substance to atleast partially reduce premature absorption of at least one of the coreingredient and the diarrheagenic substance in the gastro-intestinaltract.

The application also provides a a multiparticulate composition forpreventing or reducing potential harm from overdose ingestion,comprising: a population of particulates containing a core ingredient,the aggregate amount of the core ingredient in the population adjustedbelow a threshold dose; and a diarrheagenic coating enclosing thepopulation of the core ingredient particulates and containing an amountof a diarrheagenic substance, the amount of the diarrheagenic substancebeing sufficient to cause diarrhea to expel at least a portion of thecore ingredient from the gastro-intestinal tract of a specimen if thespecimen ingests the composition approximately at or above the thresholddose, thereby reducing potential harm from overdose ingestion of thecomposition. The multiparticulate composition may further include acontrolled-release coating containing a controlled-release substance toadjust at least one of a time and a venue of absorption of the coreingredient in the gastrointestinal tract, the controlled-release coatinginterposed between, and separating, the population of the coreingredient particulates and the diarrheagenic coating. Themultiparticulate composition may further include an enteric coatingcontaining an enteric substance, the enteric coating enclosing thediarrheagenic coating to at least partially reduce premature absorptionof at least one of the core ingredient and the diarrheagenic substancein the gastro-intestinal tract.

The application also provides a multiparticulate composition forpreventing or reducing potential harm from overdose ingestion,comprising a population of particulates, at least one of theparticulates containing: an amount of a core ingredient, the amount ofthe core ingredient being sufficiently low such that an aggregate amountof the core ingredient in the particulate population is below athreshold dose; and a diarrheagenic coating enclosing the coreingredient and containing an amount of a diarrheagenic substance, theamount of the diarrheagenic substance being sufficient to cause diarrheato expel at least a portion of the core ingredient from thegastrointestinal tract of a specimen if the specimen ingests thecomposition approximately at or above the threshold dose, therebyreducing potential harm from overdose ingestion of the composition. Themultiparticulate composition may further include a controlled-releasecoating containing a controlled-release substance to adjust at least oneof a time and a venue of absorption of the core ingredient in thegastrointestinal tract, the controlled-release coating interposedbetween, and separating, the core ingredient and the diarrheageniccoating. In certain embodiments, the venue of absorption includes atleast a portion of the small or large intestine. The multiparticulatecomposition may also include an enteric coating containing an entericsubstance, the enteric coating enclosing the particulate population toat least partially reduce premature absorption of at least one of thecore ingredient and the diarrheagenic substance in the gastro-intestinaltract. In certain embodiments, the enteric coating at least partiallyreduces the absorption of at least one of the core ingredient and thediarrheagenic substance in the stomach of the specimen.

The application further provides a multiparticulate composition forpreventing or reducing potential harm from overdose ingestion,comprising a population of particulates, at least one of theparticulates containing a mixture of: an amount of a core ingredient,the amount of the core ingredient adjusted so that an aggregate amountof the core ingredient in the particulate population is below athreshold dose; and an amount of a diarrheagenic substance, the amountof the diarrheagenic substance adjusted to cause diarrhea to expel atleast a portion of the core ingredient from the gastro-intestinal tractof a specimen if the specimen ingests the composition approximately ator above the threshold dose, thereby reducing potential harm fromoverdose ingestion of the composition. The composition may furtherinclude an enteric coating containing an enteric substance, the entericcoating enclosing the particulate population to at least partiallyreduce premature absorption of at least one of the core ingredient andthe diarrheagenic substance in the gastro-intestinal tract.

In various aspects of application, the compositions may include anorally-administrable dosage form. In certain embodiments, the dosageform can be a tablet, a capsule, a sprinkle, a caplet, a granule, aspheroid, an ellipsoid, a bead, a pellet, a chewing gum, a lozenge, acrystal, or any combination thereof.

The core ingredient described herein may include a pharmacologicalingredient, such as for example, a pharmaceutically active substance.Examples of the pharmaceutical ingredient include, without limitation,antidiabetic, analgesic, antiinflammatory agent, antirheumatic,antihypotensive agent, anti hypertensive agent, psychoactive drug,tranquillizer, antiemetic, muscle relaxant, glucocorticoid, agent fortreating an inflammatory bowel disease, antiallergic, antibiotic,antiepileptic, anticoagulant, antimycotic, antitussive, arteriosclerosisremedy, diuretic, protein, peptide, enzyme, enzyme inhibitor, goutremedy, hormone, hormone inhibitor, cardiac glycoside, immunotherapeuticagent, cytokine, laxative, lipid-lowering agent, migraine remedy,mineral product, otological agent, anti-Parkinson agent, thyroidtherapeutic agent, spasmolytic, platelet aggregation inhibitor, vitamin,cytostatic inhibitor, metastasis inhibitor, phytopharmaceutical,chemotherapeutic agent, dietary supplement, and amino acid.

Further features and advantages of the application will be apparent fromthe following description of illustrative embodiments and from theclaims.

BRIEF DESCRIPTION OF THE DRAWINGS

The following figures depict certain illustrative embodiments of theapplication in which like reference numerals refer to like elements.These depicted embodiments are to be understood as illustrative of theapplication and not as limiting in any way.

FIG. 1A depicts a schematic of an exemplary two-layer coating drugdesign.

FIG. 1B depicts a schematic of another exemplary two-layer coating drugdesign.

FIG. 2 depicts a schematic of an exemplary three-layer coating drugdesign according.

FIG. 3 depicts a schematic of an exemplary multiparticulate three-layercoating drug design.

FIG. 4 depicts a schematic of a multiparticulate drug design showingvarious combinations of particulate types.

FIG. 5 depicts a schematic embodiment of yet another multiparticulatedrug design.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

To provide an overall understanding, certain illustrative practices andembodiments will now be described, including a composition and methodfor inhibiting, preventing, or ameliorating complications associatedwith ingestion of a medicinal, chemical, and/or biological substance oragent. For convenience, in lieu of the terminology “a medicinal,chemical, and/or biological substance or agent,” the phrases“therapeutic substance,” “therapeutic compound,” and “pharmaceuticallyactive agent” are sometimes generically, and interchangeably, employedherein. However, it is understood that the scope of the application isnot limited to therapeutic substances alone, and that included withinthe scope of the compositions and methods described herein areembodiments wherein the material whose dosage is to be regulatedcontains a combination of at least one chemical, biological, and/orbiochemical substance or agent primarily intended for non-therapeutic,non-pharmacological, or even non-oral use.

In a typical embodiment, the therapeutic substance includes apharmacological compound, for example, a drug (including chemical drugsand biologics) for treating an underlying disease, ailment, or othermedical condition. Examples of a pharmacological compound (or apharmaceutically active agent) include, without limitation, a painreliever such as acetaminophen, an anti-inflammatory drug such asibuprofen, an anti-psychotic drug (neuroleptic) such as risperidone orclozapine, and a host of other chemical and/or biological compounds oragents (including, for example, biochemical compounds or agents).

In such therapeutic or pharmacological applications, the compositionformulated by the compositions and methods described herein typicallyincludes a dosage form suitable for oral administration. For example,and without limitation, the dosage form can be a combination of one ormore of a tablet, a capsule, a sprinkle, a caplet, a multi-particulateformulation (e.g., granules, spheroids, ellipsoids, beads, pellets, or acombination of these), a chewing gum, a lozenge, and a crystal. Thegeneric term “pill” is employed herein to generically refer to anembodiment of the dosage form. The pill can include a gelatin coating ora coating made of a gelatinous or gelatin-like substance.

In one aspect, the compositions and methods described herein reduce thetoxicity of chemical substances and/or biological agents down to safelevels by inducing diarrhea in a specimen. Characteristic aspects of thediarrhea include, but are not limited to, chronic and/or at least oneepisodic occurrence of increased stool production, for example,increased stool mass, increased water or other fluid content associatedwith the stool, or a combination thereof. Typically, diarrhea isassociated with abnormally frequent bowel movements that produceintestinal evacuations of increased—or possibly, though less typically,reduced-fluidity. Intestinal evacuations include, for example andwithout limitation, fecal matter typically discharged through the anusor other exit port and/or tubular means (for receiving and/or collectingbodily excrements) connected to an intestinal, post-intestinal, ordistal portion of the alimentary canal.

The term “specimen” or “subject” as used herein, typically refers to ahuman subject. However, it is understood that the scope of theapplication is not limited to human specimens, and that other animalshaving a digestive system that includes a stomach or stomach-like organand one or more intestines or intestine-like organs are within the scopeof the compositions and methods described herein. For example, andwithout limitation, specimens within the scope of the applicationinclude primates, cattle, other domesticated or wild mammalians, or acombination of these.

FIG. 1A depicts a schematic representation of an exemplary composition100 according to an embodiment of the application. In the embodimentdepicted in FIG. 1A, the dosage form 100 is a tablet, capsule, caplet,or another orally administrable dosage form—generically referred toherein as a pill.

In the illustrated embodiment, the inner core 101 contains at least onetherapeutic substance, for example and without limitation, apharmaceutically active compound. The core 101 is coated by, covered by,enveloped by, immersed in, encapsulated in, or otherwise embedded in alayer 102 containing at least one (typically active) diarrhea-inducingsubstance. The amount of the diarrheagenic substance used in layer 102is a function of one or more salient traits of members of the targetspecimen population (e.g., weight, height, gender, gastro-intestinal orother traits of the specimen population), potency and efficacy of thediarrheagenic substance 102, potency and/or toxicity of the therapeuticsubstance in the core 101, or other variables.

According to one practice, the pill 100 is uncoated (i.e., it has nolayer 103). According to an alternative practice, a portion (moretypically, the entirety) of an outer surface of the pill includes acoating 103 applied by any of a number of known techniques. The coatinglayer 103 performs one or more of the following functions, among others:(a) it protects either or both of the therapeutic and diarrhoeticcontents of the composition 100 (e.g., by discouraging, reducing, or insome instances, preventing, premature absorption and/or disintegrationof the contents in the body of the specimen); (b) it improves the visualor other physical attributes of the pill, for aesthetic or otherreasons; and (c) it carries one or more functional characteristics(e.g., pharmacological traits) of the pill.

In one aspect, the compositions and methods described herein discourage,substantially reduce, or in some instances prevent, absorption—in thestomach of the therapeutic composition present in the core 101 and/orthe diarrheagenic substance or agent present in the diarrhoetic layer102. According to one practice, the compositions, compositions, andmethods described herein accomplish this at least in part by employingan enteric material in the layer 103 to encapsulate thediarrhea-inducing compound 102 and the therapeutic substance 101. Theenteric material discourages, or more typically prevents, thediarrhoetic coating and the therapeutic substance 101 from beinginactivated by the acidity of the gastric environment. Layer 103 mayinclude one or more suitable hydrophilic polymers; enteric polymercoating materials; and/or suitable pH modifiers.

According to a typical practice, layer 103 serves as an effectivecontrolled release coating when ingested, contacted, or otherwiseexposed to an environmental fluid such as, for example and withoutlimitation, a gastric fluid or another dissolution medium in the GItract.

Among the characteristics of a typical enteric coating material are that(a) it passes through the stomach substantially unaltered; and (b) itbegins to disintegrate approximately upon or after reaching the smallintestine. This is possible at least in part because the entericmaterial resists the acidity of gastric fluids and does not disintegrateuntil essentially on or after entry into the environment of the smallintestine where the pH levels exceed those of the gastric fluids. In atypical embodiment, the enteric material is designed, configured,adapted, or otherwise selected to resist pH levels up to about 5.5 andto commence rapid disintegration when pH levels of its surroundingsexceed about 5.5. The enteric coating 103 can be designed todisintegrate if exposed to an environment characterized by any of anumber of pH levels, so that the diarrheagenic substance and therapeuticsubstance can be targeted for absorption at certain segments of thesmall intestine—such as the duodenum, jejunum, or ileum; or the colon.Absorption is designed to occur based on a particular order, thediarrheagenic substance being the first to disintegrate and be absorbedat approximately a first location in the small intestine, followed bythe therapeutic substance, which is absorbed at approximately a second,subsequent location in the small or large intestine.

The acidity of the internal environment of the stomach of a humanspecimen, for example, is generally a function of at least the amount ortype of food content in the stomach, and can be as low as about 1 pH(very acidic) and as high as about 4 pH. The alkalinity of the internalenvironment of the small intestine of a human specimen is typicallyabout 8 pH.

Once in the small or large intestine, the diarrhoetic coating 102dissolves and binds to appropriate receptors in the GI tract. Thediarrhea induced by the compositions, compositions, and methodsdescribed herein typically reverses fluid absorption and causes rapidexpulsion of the GI tract's solid, fluid, and/or other contents.Accordingly, this process reduces, or in some instances prevents,absorption of the therapeutically-active compound that is coated by thediarrheagenic substance. According to one practice, the diarrhoeticlayer 102 includes one or more suitable pH modifiers. A pH modifier mayserve to modify the pH of the local environment so that thedisintegration kinetics of a pH dependent enteric coating are dependentnot only on the specimen's physiology but also on the number of pillsingested. An alternative, or additional, function of a pH modifier is tomodulate the biological activity, potency, or efficacy of a diarroeticsubstance.

According to a typical practice, the amount of the diarrhoetic compoundin each dosage form is insufficient to produce diarrhea in a typicalmember of a broad specimen population (e.g., in a typical humanpatient). However, if several pills (constructed according to thecompositions and methods described herein) are ingested within asufficiently short time interval—such that the cumulative intake of thediarrhea-inducing substance exceeds a recommended amount or a thresholddosage—rapid and severe diarrhea ensues.

FIG. 1B depicts a schematic representation of another exemplarycomposition 150 according to an embodiment of the application. Layer 153includes one or more diarrhea-inducing substances substantiallyequivalent to the one or more diarrheagenic substances of layer 102depicted in FIG. 1A. According to one practice, the coating layer 152regulates the availability of the therapeutic substance 151 at least inpart by controlling the release of the therapeutic substance in the bodyof the specimen (e.g., this is variously known as a modified-release orcontrolled-release mechanism). The coating layer 152 may be designed tohave a pH-dependent disintegration property, as described above inrelation to the enteric coating 103 of FIG. 1A, for example.

Alternatively, or additionally, the compositions and methods describedherein may design one or more characteristics of layer 152 (e.g., athickness of layer 152, and/or a chemical, biological, biochemical,electrostatic, or other physical property of layer 152) to adjust thetime at which, or time interval during which, layer 152 disintegrates,thereby exposing and releasing the therapeutic substance 151 at adesired location in the small or large intestine and at an appropriatetime instance or time interval.

FIG. 2 depicts a schematic representation of another exemplarycomposition 200 according to an embodiment of the application. Thecomposition 200 includes features described above in relation to FIGS.1A-1B. In particular, composition includes a core 201 that contains thetherapeutic substance. The core 201 is coated by layer 202 which issubstantially similar to the time-delay layer 152 described above inrelation to FIG. 1B and controls the amount and timing of the release ofthe therapeutic substance in the core 201.

Layer 203 contains the diarrheagenic substance. This layer issubstantially the same as layer 102 of FIG. 1A or layer 153 of FIG. 1B.Layer 204 contains an enteric coating substantially the same as layer103 described in relation to FIG. 1A. The composition 200, therefore,incorporates various protective tiers of the embodiments of FIGS. 1A-1B.

FIG. 3 depicts a schematic representation of another exemplarycomposition 300 according to a multi-particulate embodiment of theapplication. According to various practices, each particulate is in theform of a granule, a spheroid (e.g., a microsphere), a bead, a pellet,an ellipsoid, or a microcapsule.

In FIG. 3, three therapeutic cores 301A-301C are depicted, though itshould be noted that a multi-particulate formulation may include as fewas two such cores. Each of the therapeutic cores 301A-301C is analogousto one or more of the cores 101, 151, and 201 of FIGS. 1A, 1B, and 2,respectively, and contains one therapeutic substance or a mixture of twoor more therapeutic substances. The contents or even the particulateshapes of the therapeutic cores 301A-301C may be essentially mutuallyidentical or different at least between two particulates. For example,and without limitation, core 301A may contain a first therapeuticsubstance, core 301B may contain a second therapeutic substancedifferent from the first therapeutic substance, and core 301C maycontain a third therapeutic substance different from each of the firstand second therapeutic substances. Alternatively, at least one of theparticulates 301A-301C may contain a mixture of two or more therapeuticsubstances, wherein the mixture is different from the contents of atleast one other of the particulates 301A-301C. Similar remarks apply tothe shapes of each of the particulates 301A-301C, wherein they may allbe identical (e.g., all may be spherical of the same size) or at leastone may have a shape different from others (it may be ellipsoidalwhereas the others are spherical, or it may be spherical but larger thanthe other spherical particulates). Various permutations of how one ormore therapeutic substances may be distributed among the two or moreparticulate cores (such as 301A-301C), as well as various permutationsof how the particulates are shaped or sized, are within the scope of thecompositions and methods of the application.

In the embodiment depicted in FIG. 3, layer 304 includes an entericcoating substantially similar to the coating layer 103 as described inrelation to FIG. 1A or coating layer 204 as described in relation toFIG. 2. Layer 304 coats the diarrhea-inducing substance layer 303, whichincludes one or more diarrhea inducing substances similar to layer 102of FIG. 1A or layer 203 of FIG. 2.

Coating 302 includes a time-delay or release-control substance similarto what was described in relation to layer 152 or layer 202 of FIGS. 1Band FIG. 2, respectively. According to one practice, one or more of thetherapeutic cores 301A-301C include an inert pharmaceutical bead that atleast partially defines the core 101 of FIG. 1A, core 151 of FIG. 1B, orcore 201 of FIG. 2. An example of such a bead is a Nu-Pareil® bead(registered to Sucrest Corporation), which is a dry, free-flowing,spherical product containing sugar and starch for use as a base uponwhich drugs or medications are deposited in the manufacture ofpharmaceutical preparations.

The resultant controlled-release bead or beads in cores 301A-301C (whichare typically, though not necessarily, essentially solid) may beenclosed by the coating 302 (which may be gelatinous or gelatin-like) inan amount sufficient to provide an effective controlled-release dosewhen ingested, contacted, or otherwise exposed to an environmental fluidsuch as, for example and without limitation, a gastric fluid or anotherdissolution medium in the GI tract.

The controlled-release formulation embodiment of the compositions,compositions, and methods described herein progressively releases thetherapeutically-active agent upon ingestion and/or exposure to gastricfluids and, subsequently, intestinal fluids. The controlled-releaseprofile of the formulations of the application may be altered, forexample, by varying the amount of the enteric coating material in layer302; varying relative amounts of plasticizers; incorporating additionalingredients or excipients (e.g., in layer 302); altering the method ofmanufacture; etc. The dissolution profile of the ultimate product mayalso be modified, for example, by increasing or decreasing the thicknessof a retardant in one or more of the coating layers 302 and 304 of FIG.3.

Turning to FIG. 4, a dosage form 400 is depicted which includes anothermulti-particulate formulation according to an exemplary embodiment ofthe application. As was described in relation to FIG. 3, at least oneparticulate may be in the form of a granule, a spheroid (e.g., amicrosphere), a bead, a pellet, an ellipsoid, or a microcapsule.

One or more therapeutic substances are present in the dosage form 400depicted in FIG. 4. In the particular embodiment of FIG. 4, a firstparticulate 410A is shown which includes a first therapeutic substancein a first core 401A having a first coating 402A. The coating is made atleast in part (though typically, essentially) of a first time-delay orcontrolled-release material not unlike coaling 152 of FIG. 1B, coating202 of FIG. 2, or coaling 302 of FIG. 3.

An optional, second particulate 410B includes a second therapeuticsubstance 401B coated by a second coating 402B, the second coating madeat least in part (though typically, essentially) of a second time-delayor controlled release material. According to various practices, thefirst and second cores 401A and 401B contain essentially identicaltherapeutic substances or essentially distinct therapeutic substances.Also according to various practices, the first and second coatings 402Aand 402B contain essentially identical time-delay or controlled-releasesubstances or essentially distinct time-delay or controlled releasesubstances.

Optionally, the dosage form 400 of FIG. 4 contains a particulate 410Cmade, at least in part, of a therapeutic substance 401C (and havingessentially no diarrhea-inducing coating or time-delay orcontrolled-release coating). The therapeutic substance 401C may have aconstitution (and at least one associated property) essentially distinctfrom those of each of the therapeutic substances in the first and secondcores 401A and 401B.

Alternatively, the therapeutic substance 401C may be essentiallyidentical in constitution (and in at least one associated biological,chemical, biochemical, physical, biophysical, and/or other salientcharacteristic) to those of at least one of the first and secondtherapeutic substances in the respective cores 401A and 401B.

The relative amounts of the time-delay or controlled-release compoundsin the particulates 410A and 410B may vary; the amounts need not beessentially identical. The relative sizes and/or shapes of theparticulates 410A-410C may vary; the sizes and/or shapes need not all beessentially identical.

In the embodiment depicted in FIG. 4, layer 404 includes an entericcoating substantially similar to the coating layer 103 as described inrelation to FIG. 1A, coating layer 204 as described in relation to FIG.2, or coating layer 304 as described in relation to FIG. 3. Layer 404coats the diarrhea-inducing substance in 403, which includes one or morediarrhea-inducing substances similar to layer 102 of FIG. 1 A, layer 203of FIG. 2, or layer 303 of FIG. 3.

Turning to FIG. 5, an embodiment is depicted showing variousconfiguration permutations that the therapeutic, diarrheagenic, andtime-delay or controlled-release substances, agents, or compounds mayhave in an exemplary multi-particulate composition 500. In particular,FIG. 5 shows that the therapeutic agent and the diarrheagenic substancecan be present in the same or in different, separate, or otherwisedistinct particulates. For example, FIG. 5 depicts: (a) a population ofat least one particulate 510A containing a therapeutic substance 501Abut essentially no diarrheagenic, time-delay, or controlled-releasesubstance; (b) a population of at least one particulate 510C containinga diarrheagenic substance 503C and having an optional enteric coating504C; and (c) a population of at least one particulate 510F containing amixture 505 of a therapeutic substance and a diarrheagenic substance. Anoptional enteric coating (not shown in the figure) analogous to 504C maybe applied to the therapeutic particulate 510A.

Also depicted in FIG. 5 is a population of at least one particulate 510Bcontaining a therapeutic core 501 B coated by a diarrheagenic layer 503Bbut essentially no time-delay or controlled-release substance.Particulate 510B is similar in constitution to the composition 100depicted in FIG. 1A.

The embodiment of FIG. 5 also shows a population of at least oneparticulate 510D containing a therapeutic core 501D coated by atime-delay or controlled-release layer 502D but essentially nodiarrheagenic substance. Particulate 510D is similar in constitution tothe composition 150 depicted in FIG. 1B.

FIG. 5 also depicts a population of at least one particulate 510Econtaining a therapeutic core 501E coated by a time-delay orcontrolled-release substance 502E, which is in turn coated by adiarrheagenic layer 503E. Particulate 510E is similar in constitution tothe composition 200 depicted in FIG. 2.

The coating 504 includes the inert coating discussed above in relationto the previous figures. In the embodiment depicted in FIG. 5, coating504 includes an enteric substance substantially similar to the coatinglayer 103 as described in relation to FIG. 1A, coating layer 204 asdescribed in relation to FIG. 2, coating layer 304 as described inrelation to FIG. 3, or coating layer 404 as described in relation toFIG. 4.

As is the case with the embodiments shown in the previous figures, thetherapeutic, time-delay or controlled-release, or diarrhea-inducingsubstances in one particulate may have a constitution (and at least oneassociated biological, chemical, biochemical, physical, biophysical, orother salient characteristic) essentially distinct from or essentiallyidentical to those of a counterpart substance in another particulate.For example, and without limitation, a diarrheagenic substance in afirst particulate may be essentially identical to or different from adiarrheagenic substance in a second particulate; and/or a pH dependententeric coating in a first particulate may disintegrate at a pH levelthat is at least approximately identical to or essentially differentfrom that at which an enteric coating in a second particulatedisintegrates.

According to a typical practice, the diarrheagenic substance (e.g.,Escherichia coli Heat Stable Enterotoxin (STa)) is introduced into thecomposition 500 such that the amount of the diarrheagenic substance isinsufficient to cause diarrhea when the medication is taken at anappropriate level, typically below or at about a recommended orthreshold level. However, an excessive intake of thetherapeutically-active compound causes a rapid diarrhea that expels thetoxic or harmful accumulation of the therapeutic substantially before itcauses significant injury, death, or other harm to the specimen.

The different populations can then be mixed in the desired ratios beforebeing filled into a final dosage form 500 such as a tablet, caplet,capsule, sprinkle, or a pill-like orally-administrable formulationdescribed above.

The release kinetics of the particulates may also be different such thatthe particulates containing the diarrheagenic substances release thediarrheagenic substance into the small or large intestine prior to thetherapeutic compound being released by other particulates. The timebetween the commencement of release of the diarrheagenic substance andthe therapeutic compound can vary. The compositions and methodsdescribed herein adjust the time to be sufficiently long to allow forsevere diarrhea to develop (for example, in a patient who has overdosedon a medication) and hence expel the therapeutic compound from the GItract prior to its absorption into the blood stream.

According to yet another embodiment, one or more populations ofparticulates exist that contain both the therapeutic compound and thediarrheagenic agent. According to one practice, in particles thatcontain both the diarrheagenic agent and the therapeutic compound, theparticles are coated on their outer-most surface with the diarrheagenicagent to ensure that the diarrheagenic substance is released into thesmall or large intestine prior to the therapeutic compound is released.Such populations of particles can be mixed together prior to beingfilled into a final dosage form, such as a capsule or sprinkle, or theycan be mixed with one or more populations that contain the therapeuticcompound but not the diarrheagenic agent and/or the diarrheagenic agentbut not the therapeutic compound prior to being filled into a finaldosage form such as a capsule or a sprinkle.

According to one practice, the dosage form is adapted to have a modifiedrelease property. The term “modified release,” as defined herein, refersto the release of the therapeutic compound at a rate such that theplasma concentration of the therapeutic compound within the person towhom the therapeutic compound has been administered is maintained withinan acceptable therapeutic range, that is, above a minimumtherapeutically-effective concentration, but below toxic levels, overthe period of time in which the therapeutic compound is released.

The compositions and methods described herein provide themodified-release property of the oral dosage form in any of a number ofways. For example, and without limitation, a modified-release carriercan be used that is incorporated into the matrix of the composition.Alternatively, or additionally, a modified-release coating may beapplied to a surface of the dosage form. In embodiments that employ amodified-release coating, the coating material is selected to achievethe desired in vitro release rate and, typically, is capable of forminga strong, continuous film that is smooth and elegant, and is able tosupport colorants and other coating additives. Additionally, the coatingmaterial has little or substantially no toxicity, is substantiallyinert, and/or is substantially tack-free.

While the therapeutic compound may have a modified-release property inthe small and/or large intestines, the diarrheagenic substance can bepackaged in a modified-release form or in an immediate-release form forwhen it reaches the small or large intestine. In a typical embodiment,the therapeutic compound has a coating to reduce, or in some instancessubstantially prevent, the absorption and/or degradation of thediarrheagenic substance in the stomach. In a typical embodiment, thecoating also substantially reduces, or in some instances prevents, theabsorption of the therapeutic compound in the stomach.

In another embodiment, the modified-release coating permits either pHdependent or pH-independent release of the therapeutic compound and/orthe diarrheagenic substance, for example and without limitation, whenexposed to the gastrointestinal liquids. A pH-dependent coating servesto release the therapeutic compound in desired locations of the GI tractfor example, the stomach, small intestine, or colon—providing anabsorption profile capable of inducing in the user a sustained releaseof opiate, for example, at least about 1 hour up to about 30 hours. Whena pH-independent coating is desired, the coating is designed to achieveoptimal release regardless of pH variations along the GI tract. In yetanother embodiment, a composition is formulated that releases a portionof the unit dose in one desired location of the GI tract (e.g., thestomach) and releases the remainder of the unit dose in another locationof the. GI tract (e.g., the small intestine).

The application also contemplates a composition comprising an agent thatwould deter tempering or breaking of the pharmaceutical composition tocause release of one or more pharmaceutically active agents includedtherein. Such an agent can be an irritant, such as for example, anirritant added to prevent someone from crushing a pill of anextended-release formulation and thereby destroying its extended-releasecharacteristics as described in U.S. Patent Application Publication No.20030125347. Examples of suitable local irritants may be of natural orsynthetic origin and include mustard and derivatives of mustard, forexample, allyl isothiocyanate and p-hydroxybenzyl isothiocyanate;capsaicinoids such as capsaicin, dihydrocapsaicin, nordihydrocapsaicin,homocapsaicin, and homodihydrocapsaicin; mint; aspirin; and acids suchas acids with one or more carboxyl moieties such as formic acid, aceticacid, propionic acid, butyric acid, valeric acid, caproic acid, caprilicacid, capric acid, oxalic acid, malonic acid, succinic acid, glutaricacid, adipic acid, maleic acid, fumaric acid, and citric acid. Preferredlocal irritants for use in the present application are capsaicinoidssuch as, for example, capsaicin.

Exemplary Pharmaceutically Active Agents or Substances

According to one practice, a therapeutic substance according to thecompositions, compositions, and methods described herein includes one ormore compounds suitable or otherwise used for the treatment,prophylaxis, cure, or mitigation of a disease in humans, othermammalians, or living beings having a gastro-intestinal system analogousto those of humans or other mammalians. Exemplary therapeutically-activesubstances include, without limitation, one or more of antifungalagents, antibacterial agents, antimicrobial agents, antiviral agents,spermicides, hormone agents, antitrichomonial agents, antiprotozoanagents, antimycoplasm agents, antiretroviral agents, nucleosideanalogues, reverse transcriptase inhibitors, protease inhibitors,contraceptive agents, sulfa drugs, sulfonamides, sulfones, hygieneagents, probiotic agents, vaccine agents, antibody agents, peptideagents, protein agents, polysaccharide agents, nucleic acids, plasmids,liposomes, carbohydrate polymers, transgenic bacteria, yeast,chemotherapeutic agents, steroid agents, growth-enhancing agents, libidoenhancers, androgenic substances, chitin derivatives, dietarysupplements, and mixtures and combinations thereof.

In an exemplary embodiment, acetaminophen is used as a therapeuticallyactive substance. The amount of acetaminophen included in each pill istypically about 500 milligrams (mg). According to another practice, theamount of acetaminophen included in each pill is about 100 mg. Accordingto another practice, the amount of acetaminophen included in each pillis about 200 mg. According to another practice, the amount ofacetaminophen included in each pill is about 300 mg. According toanother practice, the amount of acetaminophen included in each pill isabout 400 mg.

In an exemplary embodiment, the enteric coating is based on the use ofgelatin. One or more pH sensitive properties of a gelatin-based entericcoating can be modified using known methods. The amount of entericcoating can be modified to meet the desired release kinetics in thesmall or large intestine. According to another practice, the entericcoating includes EUDRAGIT-S 100® methods similar to those described, forexample, in U.S. Pat. No. 6,897,205.

A number of active pharmaceutical ingredients which are to be releasedin the small intestine or large intestine are suitable for use by thecompositions, compositions and methods described herein. Examples ofsuch pharmaceutical ingredients include, for example and withoutlimitation: antidiabetics, analgesics, antiinflammatory agents,antirheumatics, antihypotensive agents, antihypertensive agents,psychoactive drugs, tranquillizers, antiemetics, muscle relaxants,glucocorticoids, agents for treating an inflammatory bowel disease(e.g., ulcerative colitis, Crohn's disease, etc.), antiallergics,antibiotics, antiepileptics, anticoagulants, antimycotics, antitussives,arteriosclerosis remedies, diuretics, proteins, peptides, enzymes,enzyme inhibitors, gout remedies, hormones and inhibitors thereof,cardiac glycosides, immunotherapeutic agents and cytokines, laxatives,lipid-lowering agents, migraine remedies, mineral products, otologicals,anti-Parkinson agents, thyroid therapeutic agents, spasmolytics,platelet aggregation inhibitors, vitamins, cytostatics and metastasisinhibitors, phytopharmaceuticals, chemotherapeutic agents and aminoacids.

Examples of suitable active ingredients include but are not limited toacarbose, antigens, beta-receptor blockers, non-steroidal antirheumatia,cardiac glycosides, acetylsalicylic acid, virustatics, aclarubicin,acyclovir, cisplatin, actinomycin, alpha- and beta-sympatomimetics,(dmeprazole, allopurinol, alprostadil, prostaglandins, amantadine,ambroxol, amlodipine, methotrexate, Saminosalicylic acid, amitriptyline,amoxicillin, anastrozole, atenolol, azathioprine, balsalazide,beclomethasone, benzodiazepine, betahistine, bezafibrate, bicalutamide,diazepam and diazepam derivatives, budesonide, bufexamac, buprenorphine,methadone, calcium salts, potassium salts, magnesium salts, candesartan,carbamazepine, captopril, cefalosporins, cetirizine, chenodeoxycholicacid, ursodeoxycholic acid, theophylline and theophylline derivatives,trypsins, cimetidine, clarithromycin, clavulanic acid, clindamycin,clobutinol, clonidine, cotrimoxazole, codeine, caffeine, vitamin D andderivatives of vitamin D, colestyramine, cromoglicic acid, coumarin andcoumarin derivatives, cysteine, cytarabine, cyclophosphamide,ciclosporin, cyproterone, cytarabine, dapiprazole, desogestrel,desonide, dihydralazine, diltiazem, ergot alkaloids, dimenhydrinate,dimethyl sulphoxide, dimeticone, dipyridarnoi, domperidone anddomperidan derivatives, dopamine, doxazocin, doxorubizin, doxylamine,dapiprazole, benzodiazepines, diclofenac, glycoside antibiotics,desipramine, econazole, ACE inhibitors, enalapril, ephedrine,epinephrine, epoetin and epoetin derivatives, morphinans, calciumantagonists, irinotecan, modafinil, orlistat, peptide antibiotics,phenytoin, riluzoles, risedronate, sildenafil, topiramate, macrolideantibiotics, oestrogen and oestrogen derivatives, progestogen andprogestogen derivatives, testosterone and testosterone derivatives,androgen and androgen derivatives, ethenzamide, etofenamate, etofibrate,fenofibrate, etofylline, etoposide, famciclovir, famotidine, felodipine,fenofibrate, fentanyl, fenticonazole, gyrase inhibitors, fluconazole,fludarabine, fluarizine, fluorouracil, fluoxetine, flurbiprofen,ibuprofen, flutamide, fluvastatin, follitropin, formoterol, fosfomicin,furosemide, fusidic acid, gallopamil, ganciclovir, gemfibrozil,gentamicin, ginkgo, Saint John's wort, glibenclamide, urea derivativesas oral antidiabetics, glucagon, glucosamine and glucosaminederivatives, glutathione, glycerol and glycerol derivatives,hypothalamus hormones, goserelin, gyrase inhibitors, guanethidine,halofantrine, haloperidol, heparin and heparin derivatives, hyaluronicacid, hydralazine, hydrochlorothiazide and hydrochlorothiazidederivatives, salicylates, hydroxyzine, idarubicin, ifosfamide,imipramine, indometacin, indoramin, insulin, interferons, iodine andiodine derivatives, isoconazole, isoprenaline, glucitol and glucitolderivatives, itraconazole, ketoconazole, ketoprofen, ketotifen,lacidipine, lansoprazole, levodopa, levomethadone, thyroid hormones,lipoic acid and lipoic acid derivatives, lisinopril, lisuride,lofepramine, lomustine, loperamide, loratadine, maprotiline,mebendazole, mebeverine, meclozine, mefenamic acid, mefloquine,meloxicam, mepindolol, meprobamate, meropenem, mesalazine, mesuximide,metamizole, metformin, methotrexate, methylphenidate, methylprednisolone, metixene, metoclopramide, metoprolol, metronidazole,mianserin, miconazole, minocycline, minoxidil, misoprostol, mitomycin,mizolastine, moexipril, morphine and morphine derivatives, eveningprimrose, nalbuphine, naloxone, tilidine, naproxen, narcotine,natamycin, neostigmine, nicergoline, nicethamide, nifedipine, niflumicacid, nimodipine, nimorazole, nimustine, nisoldipine, adrenaline andadrenaline derivatives, norfloxacin, novamine sulfone, noscapine,nystatin, ofloxacin, olanzapine, olsalazine, omeprazole, omoconazole,ondansetron, oxaceprol, oxacillin, oxiconazole, oxymetazoline,pantoprazole, paracetamol, paroxetine, penciclovir, oral penicillins,pentazocine, pentifylline, pentoxifylline, perphenazine, pethidine,plant extracts, phenazone, pheniramine, barbituric acid derivatives,phenylbutazone, phenytoin, pimozide, pindolol, piperazine, piracetam,pirenzepine, piribedil, piroxicam, pramipexole, pravastatin, prazosin,procaine, promazine, propiverine, propranolol, propyphenazone,prostaglandins, protionamide, proxyphylline, quetiapine, quinapril,quinaprilat, ramipril, ranitidine, reproterol, reserpine, ribavirin,rifampicin, risperidone, ritonavir, ropinirole, roxatidine,roxithromycin, ruscogenin, rutoside and rutoside derivatives, sabadilla,salbutamol, salmeterol, scopolamine, selegiline, sertaconazole,sertindole, sertralion, silicates, sildenafil simvastatin, sitosterol,sotalol, spaglumic acid, sparfloxacin, spectinomycin, spiramycin,spirapril, spironolactone, stavudine, streptomycin, sucralfate,sufentanil, sulbactam, sulphonamides, sulfasalazine, sulpiride,sultamicillin, sultiam, sumatriptan, suxamethonium chloride, tacrine,tacrolimus, taliolol, tamoxifen, taurolidine, tazarotene, temazepam,teniposide, tenoxicam, terazosin, terbinafine, terbutaline, terfenadine,terlipressin, tertatolol, tetracyclins, teryzoline, theobromine,theophylline, butizine, thiamazole, phenothiazines, thiotepa, tiagabine,tiapride, propionic acid derivatives, ticlopidine, timolol, tinidazole,tioconazole, tioguanine, tioxolone, tiropramide, tizanidine, tolazoline,tolbutamide, tolcapone, toinaftate, tolperisone, topotecan, torasemide,antioestrogens, tramadol, tramazoline, trandolapril, tranylcypromine,trapidil, trazodone, triamcinolone and triamcinolone derivatives,triamterene, trifluperidol, trifluridine, trimethoprim, trimipramine,tripelennamine, triprolidine, trifosfamide, tromantadine, trometamol,tropalpin, troxerutine, tulobuterol, tyramine, tyrothricin, urapidil,ursodeoxycholic acid, chenodeoxycholic acid, valaciclovir, valproicacid, vancomycin, vecuronium chloride, Viagra, venlafaxine, verapamil,vidarabine, vigabatrin, viloazine, vinblastine, vincamine, vincristine,vindesine, vinorelbine, vinpocetine, viquidil, warfarin, xantinolnicotinate, xipamide, zafirlukast, zalcitabine, zidovudine,zolmitriptan, zolpidem, zoplicone, zotipine and the like.

Exemplary Diarrheagenic Substances or Agents

The diarrhea-inducing substances referred to in the descriptions of FIG.1A-FIG. 5 typically include one or more of a bacterial toxin (e.g.,Escherichia coli Heat Stable Enterotoxin (STa)); a viral protein; aprotein and/or peptide (e.g., guanylin and uroguanylin); a laxative(e.g., aloe vera, bisacodyl, casanthranol, cascara sagrada, castor oil,dehydrocholic acid, phenolphthalein, picosulfate, senna and sennosides);and another naturally-occurring or synthetic pharmacological compoundhaving a diarrheagenic property or capable of inducing diarrhea in asubject or specimen.

Examples of enterotoxins are listed as follows:

(J. Peptide Res., 63, 2004/200-206. (The table above compares amino acidsequences of heat-stable entero-toxins elaborated by various entericbacteria. The numbers of amino acid residues are referred to Ec-STp.Invariant residues are indicated by hatched letters. In specificembodiments, a variant or mutant enterotoxin comprises one or moremutations of amino acids that are not at the positions of the invariantresidues or alternatively comprises one or more conservative mutationsof amino acids that are at the positions of the invariant residues asshown.)

In an exemplary embodiment, STa is used as the diarrhea-inducing agent.STa is a potent toxin which—depending at least in part on its level ofpurity and/or biological activity—can induce diarrhea in humans ifingested in quantities as low as about 5 micrograms (mcg) or lower.

Examples of STa amino acid sequences are as follows:

SEQ ID NO: 9 ASN-THR-PHE-TYR-CYS-CYS-GLU-LEU-CYS-CYS-ASN-PRO-ALA-CYS-ALA-GLY-CYS-TYR SEQ ID NO: 10ASN-SER-SER-ASN-TYR-CYS-CYS-GLU-LEU-CYS-CYS-ASN-PRO-ALA-CYS-THR-GLY-CYS-TYR SEQ ID NO: 11Asn-Thr-Phe-Tyr-Cys-Cys-Glu-Leu-Cys-Cys-Asn- Pro-Ala-Cys-Ala-Gly-Cys-Tyr

According to Thompson and Giannella, Infection and Immunity (1985)47:834-836, all heat-stable enterotoxins composed of 18 amino acidssequenced then from human, porcine, and bovine isolates of E. coli haveidentical primary structures, i.e., (SEQ ID NO: 11)Asn-Thr-Phe-Tyr-Cys-Cys-Glu-Leu-Cys-Cys-Asn-Pro-Ala-Cys-Ala-Gly-Cys-Tyr.Furthermore, all 18- and 19-amino-acid heat-stable enterotoxins from E.coli share an almost identical core sequence, i.e., 14 of the 15carboxy-terminal amino acid residues are identical.

As used herein, “sequence identity” (or “% identical”) means thepercentage of identical amino acid residues at corresponding positionsin two or more sequences when the sequences are aligned to maximizesequence matching, i.e., taking into account gaps and insertions.Identity can be readily calculated by known methods, including but notlimited to those described in Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988, Biocomputing:Informatics and Genome Projects, Smith, D. W., ed., Academic Press, NewYork, 1993, Computer Analysis of Sequence Data, Part I, Griffin, A. M.,and Griffin, H. G., eds., Humana Press, New Jersey, 1994, SequenceAnalysis in Molecular Biology, von Heinje, G., Academic Press, 1987, andSequence Analysis Primer, Gribskov, M. and Devereux, I., eds., MStockton Press, New York, 1991, and Carillo, H., and Lipman, D., SIAM J.Applied Math., 48: 1073 (1988). Methods to determine identity aredesigned to give the largest match between the sequences tested.Moreover, methods to determine identity are codified in publiclyavailable computer programs. Computer program methods to determineidentity between two sequences include, but are not limited to, the GCGprogram package (Devereux, J., et al., Nucleic Acids Research 12(1): 387(1984)), BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Molec.Biol. 215: 403-410 (1990) and Altschul et al. Nuc. Acids Res. 25:3389-3402 (1997)). The BLAST X program is publicly available from NCBIand other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIHBethesda, Md. 20894, Altschul, S., et al., J. Mol. Biol. 215: 403-410(1990). The well known Smith-Waterman algorithm may also be used todetermine sequence identity.

The phrase “conservative amino acid substitution” refers to grouping ofamino acids on the basis of certain common properties. A functional wayto define common properties between individual amino acids is to analyzethe normalized frequencies of amino acid changes between correspondingproteins of homologous organisms (Schulz, G. E. and R. H. Schirmer,Principles of Protein Structure, Springer-Verlag). According to suchanalyses, groups of amino acids may be defined where amino acids withina group exchange preferentially with each other, and therefore resembleeach other most in their impact on the overall protein structure(Schulz, G. E. and R. H. Schirmer, Principles of Protein Structure,Springer-Verlag). Examples of amino acid groups defined in this mannerinclude:

-   (i) a charged group, consisting of Glu and Asp, Lys, Arg and His,-   (ii) a positively-charged group, consisting of Lys, Arg and His,-   (iii) a negatively-charged group, consisting of Glu and Asp,-   (iv) an aromatic group, consisting of Phe, Tyr and Trp,-   (v) a nitrogen ring group, consisting of His and Trp,-   (vi) a large aliphatic nonpolar group, consisting of Val, Leu and    Ile,-   (vii) a slightly-polar group, consisting of Met and Cys,-   (viii) a small-residue group, consisting of Ser, Thr, Asp, Asn, Gly,    Ala, Glu, Gln and Pro,-   (ix) an aliphatic group consisting of Val, Leu, Ile, Met and Cys,    and-   (x) a small hydroxyl group consisting of Ser and Thr.

In addition to the groups presented above, each amino acid residue mayform its own group, and the group formed by an individual amino acid maybe referred to simply by the one and/or three letter abbreviation forthat amino acid commonly used in the art.

“Peptidomimetic” as used herein refers to a compound in which at least aportion of a subject peptide of the application (e.g., an enterotoxinpeptide) is modified, and the three dimensional structure of thepeptidomimetic remains substantially the same as that of thepolypeptide. Peptidomimetics may be analogues of a subject polypeptideof the application (e.g., an enterotoxin) that are, themselves,polypeptides containing one or more substitutions or other modificationswithin the subject polypeptide sequence. Alternatively, at least aportion of the subject polypeptide sequence may be replaced with anonpeptide structure, such that the three-dimensional structure of thesubject polypeptide is substantially retained. In addition, otherpeptide portions of the subject peptide may, but need not, be replacedwith a non-peptide structure. Peptidomimetics (both peptide andnon-peptidyl analogues) may have improved properties (e.g., decreasedproteolysis, increased retention or increased bioavailability).Peptidomimetics may have improved oral availability. Each peptidomimeticmay further have one or more unique additional binding elements.

The amount of STa in each pill for human consumption, therefore, shouldbe below about 5 mcg, and even lower if the several pills are to bepermitted before diarrhea is induced. Typically, the amount of thediarrheagenic substance is a function of one or more factors, such astype and features of the specimen, desired rapidity of diarrhea onset,toxicity of the diarrheagenic substance, etc. Exemplary ranges for STadosage are given in a subsequent section below.

In the small intestine, a coating of STa dissolves and binds to STareceptors in the GI tract. Typically, STa induces a rapid-onset,significant, and usually involuntary diarrhea in the GI tract.Rapid-onset diarrhea refers herein to a diarrhea that is induced inapproximately 15-30 minutes after ingestion.

In various other practices, the compositions, compositions, and methodsdescribed herein employ other diarrhea-inducing materials such as adiarrhoetic chemical (as alternatives to, or in conjunction with, thediarrheagenic substances described above). Selection of such adiarrheagenic chemical is guided at least in part by one or more of thefollowing desirable, and in some instances essential, properties: afast-clearance property to reduce or substantially prevent a build-up ofthe chemical in the body of the specimen over time; reduced, acceptable,minimal, or in some instances even negligible long-term side effects andtoxicities (other than the diarrheagenic properties); and suitabilityfor producing a rapid-onset, severe diarrhea (to prevent, orsubstantially reduce, absorption of the therapeutic compound in the GItract when taken in excess of approximately the recommended or thresholddose).

In a typical embodiment, the diarrheagenic coating includes a biologicalagent, a chemical substance, or a biochemical compound having purgativeor cathartic properties that promote rapid peristalsis in the GI tract.Examples of a chemical substance having one or more of such propertiesinclude, for example and without limitation, acetylcholine, andacetylcholine esterase inhibitors (neostigmine).

Exemplary Enteric Substances

Enteric substances as employed by the compositions and methods describedherein—and described above in relation to the figures (e.g., layer 103of FIG. 1A)—include, for example and without limitation, one or moresuitable hydrophilic polymers such as, without limitation,hydroxypropylmethyl cellulose; carbomers; polyethylene oxides;hydroxypropyl cellulose; hydroxyethyl cellulose; carboxymethylcellulose;sodium carboxymethylcellulose; carboxyvinylpolymers; polyvinyl alcohols;glucans; scleroglucans; mannans; xanthans; carboxymethylcellulose andits derivatives; methylcellulose; cellulose; crosslinkedpolyvinylpyrrolidone; carboxymethyl starch; potassiummethacrylatedivinylbenzene copolymer; hydroxypropylcyclodextrin; alpha,beta, gamma cyclodextrin or derivatives and other dextran derivatives;natural gums; seaweed extract; plant exudate; agar; agarose; algin;sodium alginate; potassium alginate; carrageenan; kappa-carrageenan;lambda-carrageenan; fucoidan, furcellaran; laminarin; bypnea; eucheuma;gum arabic; gum ghatti; gum karaya; gum tragacanth; guar gum; locustbean gum; quince psyllium; flax seed; okra gum; arabinogalactin; pectin;scleroglucan; dextran; amylose; amylopectin; dextrin; acacia; karaya;guar; a swellable mixture of agar and carboxymethyl cellulose; aswellable composition comprising methyl cellulose mixed with a sparinglycrosslinked agar; a blend of sodium alginate; and locust bean gum.

Additionally, or alternatively, the enteric coating may include one ormore enteric polymer materials such as, without limitation, celluloseacetate phthalate, cellulose acetate trimaletate, hydroxypropylmethylcellulose phthalate, polyvinyl acetate phthalate, Eudragit.®. polyacrylic acid and poly acrylate and methacrylate coatings, polyvinylacetaldiethylamino acetate, hydroxypropyl methylcellulose acetatesuccinate, cellulose acetate trimellitate, shellac; hydrogels andgel-forming materials, such as carboxyvinyl polymers, sodium alginate,sodium carmellose, calcium carmellose, sodium carboxymethyl starch,polyvinyl alcohol, hydroxyethyl cellulose, methyl cellulose, gelatin,starch and cellulose-based cross-linked polymers.

Additionally, or alternatively, the enteric coating may include one ormore suitable pH modifiers. Examples of suitable pH modifiers include,without limitation, one or more of: an organic acid such as citric acid,fumaric acid, tartaric acid, succinic acid, ascorbic acid, acetic acid,malic acid, glutaric acid and adipic acid; a salt of these or ofinorganic acids; and magnesium hydroxide, etc.

Target Population

The compositions and methods described herein can be used to formulatetherapeutic or other pharmacological compositions in humans or otheranimals. The subject can be a human or animal patient in a generalpopulation or in a particular population. For example, a subject can bea pediatric patient, a pregnant or nursing woman, an elderly patient, ora patient having another condition or disease (e.g., an addiction to thepharmaceutically active agent or a hepatic disease) that makes thepatient more vulnerable to a higher-than-desirable level of the specificpharmaceutically active agent or any pharmaceutically active agent.Applicant considers the scope of the application to also includenon-therapeutic or non-pharmacological compositions; for example,certain compositions intended for non-human consumption can beformulated by employing the compositions and methods described herein toprevent accidental ingestion by children. Moreover, the compositions andmethods described herein can be used to regulate administration ofmedical, chemical, and/or biological substances or agents to not onlyhumans, but also animal target populations.

Dosage Control

According to one illustrative practice, a diarrhoetic dose level perpill is determined using a constraint-based algorithm as follows:

-   -   Number of pills ingested to reach serious toxic or lethal        dose×Diarrhoetic dose per pill=Total amount of diarrhoetic        needed to elicit rapid diarrheal reflex.    -   Diarrhoetic dose per pill×Number of pills needed for appropriate        therapeutic dose=Total amount of diarrhoetic that a target        specimen (e.g., human) can tolerate before diarrhea is induced.    -   Solve the equations above for the value of diarrhoetic dose per        pill.

According to one practice, the diarrhoetic dose level per pill isdetermined at least in part to induce diarrhea if one or more pills areconsumed by a specimen other than an intended specimen. For example andwithout limitation, a young child (e.g., below about 5 years of age)typically has an increased diarrheal response to STa in comparison withan adult (e.g., a human specimen approximately at or above 18 years ofage) such that a specific amount of STa in a pill may be insufficient toelicit a diarrheal response in the adult, but sufficient to elicit afast, and in some instances severe, diarrheal response in the child.

Exemplary Ranges of STa in Various Embodiments

In various embodiments, the amount of STa included in each pill conformsto one, or a union of more than one, of the following ranges: less thanabout 250 nanograms (ng); less than about 500 ng; between about 250 ngand about 500 ng; less than about 750 ng; between about 500 ng and about750 ng; less than about 1000 ng (i.e., 1 mcg); between about 750 ng andabout 1 mcg; less than about 2 mcg; between about 1 mcg and about 2 mcg;less than about 3 mcg; between about 2 mcg and about 3 mcg; less thanabout 4 mcg; between about 3 mcg and about 4 mcg; less than about 5 mcg;between about 4 mcg and about 5 mcg; less than about 6 mcg; betweenabout 5 mcg and about 6 mcg; less than about 7 mcg; between about 6 mcgand about 7 mcg; less than about 8 mcg; between about 7 mcg and about 8mcg; less than about 9 mcg; between about 8 mcg and about 9 mcg; lessthan about 10 mcg; between about 9 mcg and about 10 mcg; less than about20 mcg; between about 10 mcg and about 20 mcg; less than about 30 mcg;between about 20 mcg and about 30 mcg; less than about 40 mcg; betweenabout 30 mcg and about 40 mcg; less than about 50 mcg; between about 40mcg and about 50 mcg; less than about 60 mcg; between about 50 mcg andabout 60 mcg; less than about 70 mcg; between about 60 mcg and about 70mcg; less than about 80 mcg; between about 70 mcg and about 80 mcg; lessthan about 90 mcg; between about 80 mcg and about 90 mcg; less thanabout 100 mcg; between about 90 mcg and about 100 mcg; less than about150 mcg; between about 100 mcg and about 150 mcg; less than about 200mcg; between about 150 mcg and about 200 mcg; less than about 250 mcg;between about 200 mcg and about 250 mcg; less than about 300 mcg;between about 250 mcg and about 300 mcg; less than about 350 mcg;between about 300 mcg and about 350 mcg; less than about 400 mcg;between about 350 mcg and about 400 mcg; less than about 450 mcg;between about 400 mcg and about 450 mcg; less than about 500 mcg;between about 450 mcg and about 500 mcg; less than about 550 mcg;between about 500 mcg and about 550 mcg; less than about 600 mcg;between about 550 mcg and about 600 mcg; less than about 650 mcg;between about 600 mcg and about 650 mcg; less than about 700 mcg;between about 650 mcg and about 700 mcg; less than about 750 mcg;between about 700 mcg and about 750 mcg; less than about 800 mcg;between about 750 mcg and about 800 mcg; less than about 850 mcg;between about 800 mcg and about 850 mcg; less than about 900 mcg;between about 850 mcg and about 900 mcg; less than about 950 mcg;between about 900 mcg and about 950 mcg; less than about 1 milligram(mg); between about 950 mcg and about 1 mg; less than about 5 mg;between about 1 mg and about 5 mg; less than about 10 mg; between about5 mg and about 10 mg; less than about 15 mg; between about 10 mg andabout 15 mg; less than about 20 mg; between about 15 mg and about 20 mg;less than about 50 mg; between about 20 mg and about 50 mg; less thanabout 100 mg; and between about 50 mg and about 100 mg.

Additional support for the illustrative methods, compositions, andcompositions of the present application are described in, for example,U.S. Pat. No. 6,897,205, Multi-particulate form of medicament and U.S.Pat. No. 6,902,742, Multiparticulate modified release composition.

The '205 patent describes a multiparticulate drug form suitable foruniform release of an active pharmaceutical ingredient in the smallintestine and in the large intestine, comprising at least two forms ofpellets A and B which comprise an active pharmaceutical ingredient inthe core and have different polymer coatings which determine the releaseof the active ingredient at different pH values, characterized in thatpellet form A is provided with an inner polymer coating which enablescontinuous release of active ingredient, and has an outer entericcoating which rapidly dissolves above about pH 5.5, and pellet form B isprovided with an inner polymer coating which, in the USP release test,releases less than 20% of the active ingredient at pH 6.8 in 6 hours andreleases more than 50% of the active ingredient at pH 7.2 in 6 hours.The '205 patent further describes a process for producing amultiparticulate drug form by the different pellet forms A and B beingproduced by coating active ingredient-containing cores with the statedpolymer coatings, being mixed together and being converted into amultiparticulate drug form by introduction into a capsule or compressionto a tablet unit in the presence of excipients. The '205 patent alsodescribes the use of the described pellet forms A and B in the claimedprocess for producing a multiparticulate drug form with uniform releaseof active ingredient in the pH range of 6.8 and 7.2, corresponding tothe conditions in the small and large intestine, in particular for thetreatment of Crohn's disease or ulcerative colitis.

The '742 patent discloses a multiparticulate modified releasecomposition having a first component comprising a first population ofactive ingredient-containing particles and a second component comprisinga second population of active ingredient-containing particles. Theactive ingredient contained in the first and second components can bethe same or different and active ingredient-containing particles of thesecond component are coated with a modified release coating.Alternatively or additionally, the second population of activeingredient containing particles further comprises a modified releasematrix material. Following oral delivery, the composition in operationdelivers the active ingredient or active ingredients in a pulsatilemanner. The '742 patent further provides a multiparticulate modifiedrelease composition of which the first component is an immediate release(IR) component. Further, the modified release coating applied to thesecond population of active ingredient containing particles causes a lagtime between the release of active ingredient from the first populationof active ingredient containing particles and the release of activeingredient from the second population of active ingredient containingparticles. Similarly, the presence of a modified release matrix materialin the second population of active ingredient containing particlescauses a lag time between the release of active ingredient from thefirst population of active ingredient containing particles and therelease of active ingredient from the second population of activeingredient containing particles. The duration of the lag time may bevaried by altering the composition and/or the amount of the modifiedrelease coating and/or altering the composition and/or amount ofmodified release matrix material utilized. Thus, the duration of the lagtime can be designed to mimic a desired plasma profile. Because theplasma profile produced by the multiparticulate modified releasecomposition upon administration is substantially similar to the plasmaprofile produced by the administration of two or more IR dosage formsgiven sequentially, the multiparticulate controlled release compositiondescribed in the '742 patent is particularly useful for administeringactive ingredients for which patient tolerance may be problematical.This multiparticulate modified release composition is thereforeadvantageous for reducing or minimizing the development of patienttolerance to the active ingredient in the composition.

EXAMPLES

The Examples described below involve animal models. Although Applicanthas included and will continue to include adult mice as study subjects,other animal models are also contemplated. For example, models based oncats, dogs, pigs, piglets, calves, rabbit, rats, and primates can alsobe employed for the testing described below. Animal maintenance andtreatments were and will be conducted in accordance with the NationalInstitute of Health Guide for Animal Welfare, as approved byInstitutional Animal Care and Use Committee. Further, toxicity, safety,and efficacy studies will be conducted through human clinical trials incases where the compositions of the application are intended for humanuse. Such clinical trials will be designed and conducted using protocolsapproved under applicable laws and regulations.

Example 1 Establishment of STa Dose-Response Curve of STa to EstablishDiarrheagenic Dose in a Population of Adult Mice Diarrheal ResponseAssay

0.5 ml of 10% glucose solution with STa at different dilutions wereadministered to adult male Swiss Webster mice (each weighing 27-33grams) by gavage. Mice were weighed prior to STa challenge. At eachdilution of STa, 10 mice were utilized for establishing a statisticalcharacterization of the diarrheal response (mean and standarddeviation). After STa challenge, at approximately 90 minutes, theanimals were sacrificed and the gut is removed and weighed. Theremaining carcass is also weighed. The gut/body weight ratio isestablished as the in-vivo assay of quantifying the diarrheal responseto exogenous STa.

Results are shown below:

Mouse Group Mean (Gut/Body Standard (10 Mice in each Group) Weight)Ratio Deviation Control (Saline) .1084 .0066  10 MU of STa .1066 .0094100 MU of STa .1144 .0065 1000 MU of STa  .1339 .0047 ***One MU is theamount of toxin which produces an intestinal weight/carcass weight ratioof ≧0.083 in 3-day-old mice.

Giannella describes a suckling mouse model for detection of aheat-stable E. coli enterotoxin, Infection and Immunity (1976) 14:95-99.

Characterization of the Diarrheal Dose Response Curve

In a pharmaceutical composition including the STa peptide with atherapeutically active compound, the amount of STa peptide released froma pill, tablet, capsule, or any oral pharmaceutical composition shouldnot elicit a diarrheal response in a human or animal if the dose of thepharmaceutical composition is approximately equal to the recommendedtherapeutic dose of the composition. However, the cumulative amount ofSTa peptide should be enough to elicit a diarrhea) response in a humanor animal if the pharmaceutical composition is administered in amountssignificantly exceeding the therapeutic dose and approaching quantitiesthat would be toxic, or lethal, or otherwise harmful to a human oranimal.

Example 2 Dose-Response Curve for Acetaminophen-Induced Hepatotoxicity

Data from several studies suggests that acetaminophen is responsible for39 percent of all cases of acute liver failure in the United States.E.g., Lee, New England J. Med. (2003) 349: 474-485, Shankar et al.,Toxicol. Sci. (2003) 73: 220-234. Acetaminophen-induced hepatotoxicityis also a well-established model of fulminant hepatic failure. E.g.,Newsome et al., Liver Transplantation (2000) 6: 21-31.

Acetaminophen Administration

Food, but not water, will be removed 12 hours before acetaminophenadministration. The method for acetaminophen administration will be asdescribed by Walker et al, Lab Invest. (1980) 42:181-189. Briefly,acetaminophen will be dissolved in warm distilled water at variousdilutions and given to mice by gavage at a volume of 0.5 ml. At eachdilution of acetaminophen, 5 mice will be utilized for statisticalcharacterization (mean and standard deviation) of physiologic responsesto acetaminophen challenges.

Dilutions of Acetaminophen to be administered (mg/kg):

0 5 20 40 80 100 200 500 750 1000

At 6 hours post acetaminophen challenge, blood samples for biochemicalassays will be collected under diethyl ether anesthesia.

Histopathology

At 6 hours post acetaminophen challenge (and after blood samples havebeen collected), livers will be surgically removed from mice underdiethyl ether anesthesia. Portions of liver will be taken from the leftlateral lobes and washed with ice-cold normal saline (0.9% NaCl), cutinto small pieces, and fixed immediately in 10% phosphate-bufferedformalin for 48 h. The liver tissue will then be transferred to 70%ethyl alcohol and stored until processed. The liver specimens will beprocessed, embedded in paraffin, sectioned at 5 μm, and stained withhematoxylin and eosin (H&E) for histological examination under a lightmicroscope. The extent of liver injury will be estimated by certifiedpathologists from the animal disease diagnostic laboratory of MichiganState University.

Biochemistry Assays

From the collected blood samples, plasma alanine aminotransferase (ALT)will be determined as a biochemical marker of liver damage. The plasmaacetaminophen levels will also be determined using standard assays(Walker et al., supra).

Example 3 Evaluation of Protective Drug Delivery Model to PreventAcetaminophen-Induced Hepatic Failure in Adult Mice

In this section, a formulation of acetaminophen combined with STa willbe evaluated for preventing hepatotoxicity due to acute acetaminophenoverdosing. There will be three experimental groups and one controlgroup. Group A will include mice administered a hepatotoxic dose ofacetaminophen in a particulate (solid) form after a diarrheagenic doseof STa. Group B will include mice administered a hepatotoxic dose ofacetaminophen dissolved in solution after a diarrheagenic dose of STa.Group C will include mice administered a therapeutic dose ofacetaminophen in particulate (solid) form after a non-diarrheagenic doseof STa. The control group will consist of mice administered ahepatotoxic dose of acetaminophen in particulate (solid) form withoutany STa administration. Each experimental group and the control groupwill consist of 5 mice. Mice will be weighed at the beginning of theexperiment and six hours after acetaminophen challenge. During thisinterval, mice will not be afforded water or solid nutrition.

Administration of STa to Experiment Groups

As determined in section 1, a diarrheagenic dose of STa will beadministered by gavage to mice from groups A and B 30 minutes prior toacetaminophen challenge. Group C mice will be administered anon-diarrheagenic dose of STa (approximately 5% of the dose of adiarrheagenic response as determined in section 1). The control micewill not be administered any STa (only 0.5 ml of 10% glucose) 30 minutesprior to acetaminophen challenge.

Acetaminophen Administration

30 minutes after STa administration, mice from group A will beadministered a hepatotoxic dose of acetaminophen prepared in solidformulation (15-20 mg), by gavage.

Thirty minutes after ST administration, mice from group B will beadministered a hepatotoxic dose of acetaminophen in 0.5 ml bolussolution. The acetaminophen solution will be prepared by dissolvingacetaminophen in warm distilled water. The concentration will bedetermined from the results of section 2.

In the group C mice, a therapeutic dose of acetaminophen (approximately10 mg/kg) will be administered using a 0.5 ml bolus solution 30 minutesafter a non-diarrheagenic STa challenge. Approximately 90 minutes afteracetaminophen administration, blood samples will be drawn under diethylether anesthesia. Then, mice will be sacrificed under diethyl etheranesthesia and liver tissue samples will be obtained as described in thepreceding section.

Control mice will be given a solid formulation of acetaminophen at ahepatotoxic dose. Control mice will not be administered any STa prior tothe acetaminophen challenge.

Biochemical Assays and Histopathology

In the experimental groups A and B, and the control group, the mice willbe sacrificed 6 hours after acetaminophen challenge under diethyl etheranesthesia. Prior to sacrifice, blood samples will be drawn underanesthesia to measure ALT and acetaminophen levels in plasma. Liversamples will be obtained and processed and submitted for histopathology.

Results and Analysis of Data

The plasma ALT levels in experimental groups A, B, C, and the controlgroups will be compared to determine whether the STa treatment alongwith the toxic levels of acetaminophen resulted in a significantreduction of the hepatotoxicity as measured by normal or near normalserum ALT levels. Histopathology to visually identify necrosis andevidence of acute liver injury will be a second means of assessingacetaminophen effects with and without prior STa administration. Thedifference in mice weights at the beginning and end of the experiments(6 hour duration) will be considered as a surrogate for the magnitude ofthe secretory diarrheal response. The acetaminophen plasma levels willalso be measured in all groups to investigate whether STa-inducedsecretory diarrhea significantly reduces the bioavailability ofacetaminophen.

INCORPORATION BY REFERENCE

All references including scientific literature and patent literaturecited herein are hereby incorporated by reference in their entirety.

EQUIVALENTS

Many other equivalents to the specific embodiments of the applicationand the specific aspects and practices associated with the compositionsand methods described herein exist. Accordingly, the application is notto be limited to the embodiments, aspects, practices, and methodsdisclosed herein, but it is to be understood from the following claims,which are to be interpreted as broadly as allowed under the law.

1-21. (canceled)
 22. A therapeutic composition comprising: apharmaceutically active agent; and a diarrheagenic agent that is anenterotoxin comprising an amino acid sequence that is 95% identical toSEQ ID NO: 10; and wherein said diarrheagenic agent induces diarrhea toreduce the harmful or undesired side effects of the pharmaceuticallyactive agent when the pharmaceutically active agent is ingested at adose higher than the prescribed dose.
 23. The therapeutic composition ofclaim 22, wherein the diarrhea reduces the amount of thepharmaceutically active agent absorbed by the subject.
 24. Thetherapeutic composition of claim 22, wherein the therapeutic compositionis formulated such that ingestion by a subject of the therapeuticcomposition at or lower than the prescribed dosage does not causediarrhea in the subject.
 25. The therapeutic composition of claim 22,wherein the diarrheagenic agent is coated onto the pharmaceuticallyactive agent.
 26. The therapeutic composition of claim 22, furthercomprising an additional coating.
 27. The therapeutic composition ofclaim 26, wherein the additional coating comprises an enteric coating.28. The therapeutic composition of claim 22, wherein the therapeuticcomposition comprises an orally administrable dosage form.
 29. Thetherapeutic composition of claim 28, wherein orally administrable dosageform is a tablet.
 30. A therapeutic composition comprising: apharmaceutically active agent; and a diarrheagenic agent; wherein thediarrheagenic agent is an enterotoxin comprising an amino acid sequenceat least 95% identical to SEQ ID NO: 10; and said therapeuticcomposition comprises less than about 1 mg of the enterotoxin.
 31. Amethod for reducing harmful or undesired side effects in a subject,caused by ingestion of a pharmaceutically active agent at a dose higherthan the prescribed dosage comprising: administering to the subject atherapeutic composition comprising: a) a pharmaceutically active agent;and b) a diarrheagenic agent comprising an amino acid sequence at least95% identical to SEQ ID NO: 10; wherein said diarrheagenic agent inducesdiarrhea to reduce the harmful or undesired side effects of thepharmaceutically active agent when the pharmaceutically active agent isingested at a dose higher than the prescribed dose.
 32. The therapeuticcomposition of claim 22 or 30, wherein the pharmaceutically active agentis selected from antidiabetic, analgesic, antiinflammatory agent,antirheumatic, antihypotensive agent, anti hypertensive agent,psychoactive drug, tranquillizer, antiemetic, muscle relaxant,glucocorticoid, agent for treating an inflammatory bowel disease,antiallergic, antibiotic, antiepileptic, anticoagulant, antimycotic,antitussive, arteriosclerosis remedy, diuretic, protein, peptide,enzyme, enzyme inhibitor, gout remedy, hormone, hormone inhibitor,cardiac glycoside, immunotherapeutic agent, cytokine, laxative,lipid-lowering agent, migraine remedy, mineral product, otologicalagent, anti-Parkinson agent, thyroid therapeutic agent, spasmolytic,platelet aggregation inhibitor, vitamin, cytostatic inhibitor,metastasis inhibitor, phytopharmaceutical, chemotherapeutic agent, aminoacid(s), and any combination thereof.
 33. The method of claim 31,wherein the pharmaceutically active agent is selected from antidiabetic,analgesic, antiinflammatory agent, antirheumatic, antihypotensive agent,anti hypertensive agent, psychoactive drug, tranquillizer, antiemetic,muscle relaxant, glucocorticoid, agent for treating an inflammatorybowel disease, antiallergic, antibiotic, antiepileptic, anticoagulant,antimycotic, antitussive, arteriosclerosis remedy, diuretic, protein,peptide, enzyme, enzyme inhibitor, gout remedy, hormone, hormoneinhibitor, cardiac glycoside, immunotherapeutic agent, cytokine,laxative, lipid-lowering agent, migraine remedy, mineral product,otological agent, anti-Parkinson agent, thyroid therapeutic agent,spasmolytic, platelet aggregation inhibitor, vitamin, cytostaticinhibitor, metastasis inhibitor, phytopharmaceutical, chemotherapeuticagent, amino acid(s), and any combination thereof.